Selection of Chloroplasts by Laser Microbeam Microdissection for Single-Chloroplast PCR

Meimberg, H.; Thalhammer, S.; Brachmann, Andreas ; Müller, B.; Eichacker, L.A.; Heckl, W.M. und Heubl, G. (Juni 2003): Selection of Chloroplasts by Laser Microbeam Microdissection for Single-Chloroplast PCR. In: BioTechniques, Bd. 34, Nr. 6: S. 1238-1243

Abstract

Laser microbeam microdissection and laser pressure catapulting offer the possibility of separating cell compartments, thus allowing for contamination-free analysis. Using these methods, we were able to select single chloroplasts of Nicotiana tabacum. Starting from homogenized leaf material, chloroplasts were purified by differential centrifugation and applied directly onto a poly-ethylene-naphthalate membrane that was mounted on a microscope slide. Single chloroplasts were dissected under microscopic control and catapulted into a PCR tube. Subsequent PCR of a spacer region between the trnT and trnF genes verified the successful amplification of DNA from a single chloroplast. The advantage of this method compared to the use of capillaries or optical tweezers is that one is able to prepare high numbers of samples in a short time.

• Dokumententyp: Zeitschriftenartikel
• Publikationsform: Publisher's Version
• Fakultät: Biologie > Department Biologie I
• Themengebiete: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
• Sprache: Englisch